Field of Invention
The invention pertains to dsRNA particles, recombinant RNA and DNA molecules, and methods of production and use and the expression of proteins therefrom.
Background Information
Engineered viral systems present great opportunities for therapeutic and commercial applications. The genomes of many viruses have been sequenced and characterized with respect the replication, packaging, immune evasion, protective antigens, killer toxin, immunity proteins, etc. Utilizing this information, viruses have been altered for use as attenuated vaccines or engineered for use as protein expression systems for use in gene therapy, vaccines and protein products. Examples of viruses that have been used in this manner include alphaviruses, adenoviruses, baculoviruses, pox viruses, rhabdoviruses, picornaviruses, noroviruses, niedoviruses, nidoviruses, and flaviviruses.
Various methods are available for the production of proteins. Standard method of producing proteins include transformation of cells and genomic insertion of DNA by homologous or non-homologous recombination. Episomal plasmids, end joining, and artificial chromosomes or phages have also been used to get DNA into cells and to achieve transcription and translation of proteins. All of these methods are susceptible to gene silencing by epigenetic factors or by the location in the genome where the DNA happens to insert. All of these methods require the initiation of transcription and therefore positional effects in the genome, promoter strength, and gene regulation all influence their extent of expression.
It would therefore be advantageous to be able to engineer and utilize dsRNA viruses naturally found in a wide variety of yeast or fungi to deliver and/or propagate a recombinant dsRNA molecule in a virus or particle, which could be introduced into an organism to be treated to regulate gene expression in the organism or in a pathogen infecting the organism. It would also be advantageous to be able to use such systems for the production of desirable proteins or for other industrial application, such as for controlling the growth of unwanted organisms, or in the production of therapeutic agents. With such compositions and methods, a highly useful system for efficient production and delivery of packaged dsRNA could be achieved.